RNA-Seq of mouse liver transcriptome: role of genetic sex and hepatic ERa

The study was aimed at evaluating the extent of sex differences in adult mouse liver and the role of the hepatic estrogen receptor (ER) in determining such a difference. Overall design: All experiments were done with littermates of 8 months of age. Prior euthanasia, mice were fasted 6h to avoid diet-induced activation of the hepatic ERα. RNA was isolated from homogenates of livers from intact male and female mice. As the phase of the estrous cycle impacts on liver transcriptome, the comparative study was carried out using females in two phases of the estrous cycle characterized by high and low circulating estrogens (proestrus, P and metestrus, M, respectively). To verify the impact of hepatic ERa on liver transcriptome, RNA-seq analysis was also done on control (ERa floxed) or LERKO (liver ERa knockout) mice. Illumina TruSeq RNA Sample Prep V2 was used for library construction and samples were sequenced on Illumina NextSeq, producing 2X75bp paired end reads.